Virtual Screening of Kelch-like ECH-Associated Protein 1−Nuclear Factor Erythroid 2-Related Factor 2 (Keap1−Nrf2) Inhibitors and In Vitro Validation

Molecules, Год журнала: 2025, Номер 30(8), С. 1815 - 1815

Опубликована: Апрель 17, 2025

The transcription factor erythroid 2-related 2 (Nrf2) is a central regulator of cellular defense mechanisms against oxidative stress and inflammation. Keap1 (Kelch-like ECH-associated protein 1) regulates Nrf2 activity by ubiquitination-mediated cytoplasmic retention, thereby suppressing its nuclear translocation subsequent transcriptional activation genes encoding phase II detoxifying enzymes. Using structure-based virtual screening approach, we screened ~16,000 natural compounds to identify Keap1-Nrf2 PPI inhibitors. Nine were identified based on their high binding affinities favorable interactions with Keap1, primarily through non-covalent interactions. To validate the stability these inhibitors, molecular dynamics (MD) simulations performed, confirming robustness Keap1-inhibitor complexes over time. Subsequent in vitro assays human epithelial keratinocyte cells (HaCaT) revealed that six notably upregulated mRNA expression, regis tering increases from 23% 50% comparison control. Notably, chebulinic acid emerged as most potent compound, demonstrating greatest elevation expression. Penetration studies further showed acid, when encapsulated silk fibroin, achieved 0.14% penetration rate after 24 h though it could not penetrate into stratum corneum alone. This result highlighted potential use anti-aging skincare formulations. Collectively, our findings affirmed docking reliable effective approach for identification novel agents targeting pathway.

Язык: Английский

Cdc48 plays a crucial role in redox homeostasis through dynamic reshaping of its interactome during early stationary phase.

Redox Biology, Год журнала: 2025, Номер unknown, С. 103651 - 103651

Опубликована: Май 1, 2025

Most microbial cells on earth predominantly exist in non-proliferating, dormant conditions, such as the stationary state. The phase is a crucial stage during cellular lifespan, which requires homeostatic rewiring for long-term viability and rapid responses to environmental changes. Here, we show that entry yeast accompanied by increased cytosolic mitochondrial oxidation, imposing stress proteostasis network. We establish functional link between redox protein homeostasis, mediated key quality control member, Cdc48/p97/VCP. Comparative proteomic analysis of post-mitotic reveals while global proteome remains largely stable first stages phase, Cdc48 interactome undergoes significant remodeling, including altered interactions with antioxidants its cofactors Shp1/Ubx1 Ubx2. To challenge Cdc48's capacity redox-switch early utilized proteomics map changes reversible oxidation modification cysteines upon phase. revealed temporal Cdc48-Cys115 regulatory event essential stationary-phase survival modulation. Cys115-to-serine mutation significantly reduced longevity oxidative sensitivity, correlating disrupted antioxidants, cofactor Shp1, specifically phosphorylated form Shp1. Taken together, these findings identify new thiol switch degradation pathway, further defining novel roles reshaping

Язык: Английский